Loading libraries

dyn.load("/home/moyra.lawrence/miniconda3/envs/monocle3/lib/libgeos_c.so.1")
library(Seurat)
## Warning in fun(libname, pkgname): rgeos: versions of GEOS runtime 3.11.0-CAPI-1.17.0
## and GEOS at installation 3.8.0-CAPI-1.13.1differ
library(patchwork)
library(ggplot2)
library(gridExtra)
library(openxlsx)
library(svglite)
library(dplyr)
library(gt)
library(tidyverse)
library(stringi)
library(SeuratObject)
library(biomaRt)
library(stringr)

#Loading Hu et al., 2023

Hu <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Hu')
Hu = CreateSeuratObject(counts = Hu, project = "Hu")

Checking filtering

Hu[["percent.mt"]] <- PercentageFeatureSet(Hu, pattern = "^mt-")
VlnPlot(Hu, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(Hu, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(Hu, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(Hu, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

##Filtering

1000 < nFeature_RNA
2000 < nCount_RNA <100,000
0 =< percent.mt < 10

Hu.filtered <- subset(Hu, subset = nFeature_RNA > 1000 & nCount_RNA > 2000 & nCount_RNA < 100000 & percent.mt < 10)
Hu.filtered
## An object of class Seurat 
## 32285 features across 9795 samples within 1 assay 
## Active assay: RNA (32285 features, 0 variable features)

Confirming filtering

vp1 <- VlnPlot(Hu.filtered, pt.size=0, features = c("nFeature_RNA")) +  theme(legend.position = 'none')
vp2 <- VlnPlot(Hu.filtered, pt.size=0, features = c("nCount_RNA")) +  theme(legend.position = 'none')
vp3 <- VlnPlot(Hu.filtered, pt.size=0, features = c("percent.mt")) + theme(legend.position = 'none')
vp1.log <- VlnPlot(Hu.filtered, pt.size=0, features = c("nFeature_RNA"),log=TRUE)
vp2.log <- VlnPlot(Hu.filtered, pt.size=0, features = c("nCount_RNA"),log=TRUE)
vp3.log <- VlnPlot(Hu.filtered, pt.size=0, features = c("percent.mt"),log=TRUE) 
grid.arrange(vp1,vp2,vp3, nrow=1)
grid.arrange(vp1.log,vp2.log,vp3.log, nrow=1)
FeatureScatter(Hu.filtered, "nCount_RNA", "percent.mt", pt.size=0.5) + scale_y_continuous(limits=c(0,100)) + scale_x_continuous(limits=c(0,100000))
FeatureScatter(Hu.filtered, "nCount_RNA", "nFeature_RNA", pt.size=0.5) + scale_y_continuous(limits=c(0,12000)) + scale_x_continuous(limits=c(0,100000))

#Loading Xu et al., 2022

XuESC1 <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_ES1/ES1')
XuESC2 <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_ES2/ES2')
XuEPS1 <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_EPS1/EPS1')
XuEPS2 <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_EPS2/EPS2')
XuEPSTPS <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_EPS_TPS/EPS-TPS2')
Xu2CTPS <- Read10X(data.dir = '/home/moyra.lawrence/R_analysis/Comparison/Xu/Xu_2C_TPS/2C-TPS1')
XuESC1 = CreateSeuratObject(counts = XuESC1, project = "XuESC")
XuESC2 = CreateSeuratObject(counts = XuESC2, project = "XuESC")
XuEPS1 = CreateSeuratObject(counts = XuEPS1, project = "XuEPSC")
XuEPS2 = CreateSeuratObject(counts = XuEPS2, project = "XuEPSC")
XuEPSTPS = CreateSeuratObject(counts = XuEPSTPS, project = "XuEPSTPS")
Xu2CTPS = CreateSeuratObject(counts = Xu2CTPS, project = "Xu2CTPS")

Quality plot XuESC1

XuESC1[["percent.mt"]] <- PercentageFeatureSet(XuESC1, pattern = "^mt-")
VlnPlot(XuESC1, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(XuESC1, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(XuESC1, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(XuESC1, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

### Quality plot XuESC2

XuESC2[["percent.mt"]] <- PercentageFeatureSet(XuESC2, pattern = "^mt-")
VlnPlot(XuESC2, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(XuESC2, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(XuESC2, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(XuESC2, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

### Quality plot XuEPS1

XuEPS1[["percent.mt"]] <- PercentageFeatureSet(XuEPS1, pattern = "^mt-")
VlnPlot(XuEPS1, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(XuEPS1, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(XuEPS1, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(XuEPS1, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

### Quality plot XuEPS2

XuEPS2[["percent.mt"]] <- PercentageFeatureSet(XuEPS2, pattern = "^mt-")
VlnPlot(XuEPS2, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(XuEPS2, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(XuEPS2, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(XuEPS2, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

### Quality plot XuEPSTPS

XuEPSTPS[["percent.mt"]] <- PercentageFeatureSet(XuEPSTPS, pattern = "^mt-")
VlnPlot(XuEPSTPS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(XuEPSTPS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(XuEPSTPS, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(XuEPSTPS, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

Quality plot Xu2CTPS

Xu2CTPS[["percent.mt"]] <- PercentageFeatureSet(Xu2CTPS, pattern = "^mt-")
VlnPlot(Xu2CTPS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(Xu2CTPS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(Xu2CTPS, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(Xu2CTPS, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

##Filtering Xu

2000 < nFeature_RNA
4000 < nCount_RNA <100,000
0 =< percent.mt < 10 (12 for EPS2, 15 for 2CTPS)

XuESC1.filtered <- subset(XuESC1, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 10)
XuESC2.filtered <- subset(XuESC2, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 10)
XuEPS1.filtered <- subset(XuEPS1, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 10)
XuEPS2.filtered <- subset(XuEPS2, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 10)
XuEPSTPS.filtered <- subset(XuEPSTPS, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 12)
Xu2CTPS.filtered <- subset(Xu2CTPS, subset = nFeature_RNA > 2000 & nCount_RNA > 4000 & nCount_RNA < 100000 & percent.mt < 15)

Confirming filtering

VlnPlot(XuESC1.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(XuESC2.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(XuEPS1.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(XuEPS2.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(XuEPSTPS.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(Xu2CTPS.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

#Cossec

countsData <- read.delim(file="/home/moyra.lawrence/R_analysis/Comparison/Cossec/GSE180679_counts_run1.csv.gz", header=TRUE, sep=",", row.names=1)
metaData <- read.delim(file="/home/moyra.lawrence/R_analysis/Comparison/Cossec/GSE180679_meta_run1.csv.gz", header=TRUE, sep=",", row.names=1)


gene2 <- rownames(countsData)
gene3 <- str_replace(gene2,
                    pattern = ".[0-9]+$",
                    replacement = "")
ensembl = useMart(biomart = "ensembl", dataset="mmusculus_gene_ensembl")
ens2 = getBM(attributes=c('ensembl_gene_id', 'mgi_symbol'), 
              filters = 'ensembl_gene_id', 
              values = gene3, 
              mart = ensembl)
countsData$Gene <- gene3

merge2 <- merge(countsData, ens2, by.x="Gene", by.y = "ensembl_gene_id")
merge2["mgi_symbol"][merge2["mgi_symbol"] == ''] <- "Unknown"
names2 <- make.unique(merge2$mgi_symbol)
names2 <- gsub("_", "-", names2)
names2 <- gsub("\\.", "-", names2)
names2 <- make.unique(names2)
rownames(merge2) <- names2
merge2 <- merge2[,-1] 
merge2 <- merge2[,-5377]

Cossec <- CreateSeuratObject(counts=merge2, project="Cossec", meta.data=metaData)
Cossec$sample <- NULL
Cossec$index <- NULL
Cossec$well_coordinates<- NULL
Cossec$FSC.A<- NULL
Cossec$FSC.W<- NULL
Cossec$FSC.H<- NULL
Cossec$SSC.A<- NULL
Cossec$SSC.W<- NULL
Cossec$SSC.H<- NULL
Cossec$PE.A<- NULL
Cossec$PE.Texas.Red.A<- NULL
Cossec$PE.Cy5.A<- NULL
Cossec$PE.Cy5.5.A<- NULL
Cossec$PE.Cy7.A<- NULL
Cossec$APC.A<- NULL
Cossec$Alexa.Fluor.700.A<- NULL
Cossec$APC.Cy7.A <- NULL
Cossec$FITC.A <- NULL
Cossec$PerCP.Cy5.5.A <- NULL
Cossec$BV421.A <- NULL
Cossec$AmCyan.A <- NULL
Cossec$BV605.A <- NULL
Cossec$BV650.A <- NULL
Cossec$BV711.A <- NULL
Cossec$BV786.A <- NULL
Cossec$Time <- NULL
Cossec$XLoc <- NULL
Cossec$YLoc <- NULL

Cossec$orig.ident <- "Cossec"

Quality plot Cossec

Cossec[["percent.mt"]] <- PercentageFeatureSet(Cossec, pattern = "^mt-")
VlnPlot(Cossec, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
VlnPlot(Cossec, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)
FeatureScatter(Cossec, feature1 = "nCount_RNA", feature2 = "percent.mt")
FeatureScatter(Cossec, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")

##Filtering Cossec

2000 < nFeature_RNA
2000 < nCount_RNA <100,000

Cossec.filtered <- subset(Cossec, subset = nFeature_RNA > 2000 & nCount_RNA > 2000 & nCount_RNA < 100000)
Cossec.filtered
## An object of class Seurat 
## 52776 features across 2458 samples within 1 assay 
## Active assay: RNA (52776 features, 0 variable features)

Confirming filtering

vp1 <- VlnPlot(Cossec.filtered, pt.size=0, features = c("nFeature_RNA")) +  theme(legend.position = 'none')
vp2 <- VlnPlot(Cossec.filtered, pt.size=0, features = c("nCount_RNA")) +  theme(legend.position = 'none')
vp1.log <- VlnPlot(Cossec.filtered, pt.size=0, features = c("nFeature_RNA"),log=TRUE)
vp2.log <- VlnPlot(Cossec.filtered, pt.size=0, features = c("nCount_RNA"),log=TRUE)
grid.arrange(vp1,vp2, nrow=1)
grid.arrange(vp1.log,vp2.log, nrow=1)
FeatureScatter(Cossec.filtered, "nCount_RNA", "nFeature_RNA", pt.size=0.5) + scale_y_continuous(limits=c(0,12000)) + scale_x_continuous(limits=c(0,100000))

#Yu

#listAttributes(ensembl, page="feature_page")
YuC <- data.table::fread("/home/moyra.lawrence/R_analysis/Comparison/Yu/GSE166027_single_cell_gene_expression_Control(1).csv.gz", data.table = FALSE)
names(YuC)[2] <- 'Gene'
YuC <- YuC[,-1]
YuC <- YuC[,-2]
YuC <- YuC[,-2]
YuC <- YuC[,-2]
gene2 <- YuC$Gene
gene3 <- str_replace(gene2,
                    pattern = ".[0-9]+$",
                    replacement = "")
ensembl = useMart(biomart = "ensembl", dataset="mmusculus_gene_ensembl")
ens2 = getBM(attributes=c('ensembl_gene_id', 'mgi_symbol'), 
              filters = 'ensembl_gene_id', 
              values = gene3, 
              mart = ensembl)
YuC$Gene <- gene3

merge2 <- merge(YuC, ens2, by.x="Gene", by.y = "ensembl_gene_id")
merge2["mgi_symbol"][merge2["mgi_symbol"] == ''] <- "Unknown"
names2 <- make.unique(merge2$mgi_symbol)
names2 <- gsub("_", "-", names2)
names2 <- gsub("\\.", "-", names2)
names2 <- make.unique(names2)
rownames(merge2) <- names2
merge2 <- merge2[,-1] 
merge2 <- merge2[,-3839]

YuC <- CreateSeuratObject(counts=merge2, project="YuC")
YuC$orig.ident
## bc0001 bc0002 bc0003 bc0004 bc0005 bc0006 bc0007 bc0008 bc0009 bc0010 bc0011 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0012 bc0013 bc0014 bc0015 bc0016 bc0017 bc0018 bc0019 bc0020 bc0021 bc0022 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0023 bc0024 bc0025 bc0026 bc0027 bc0028 bc0029 bc0030 bc0031 bc0032 bc0033 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0034 bc0035 bc0036 bc0037 bc0038 bc0039 bc0040 bc0041 bc0042 bc0043 bc0044 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0045 bc0046 bc0047 bc0048 bc0049 bc0050 bc0051 bc0052 bc0053 bc0054 bc0055 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0056 bc0057 bc0058 bc0059 bc0060 bc0061 bc0062 bc0063 bc0064 bc0065 bc0066 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0067 bc0068 bc0069 bc0070 bc0071 bc0072 bc0073 bc0074 bc0075 bc0076 bc0077 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0078 bc0079 bc0080 bc0081 bc0082 bc0083 bc0084 bc0085 bc0086 bc0087 bc0088 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0089 bc0090 bc0091 bc0092 bc0093 bc0094 bc0095 bc0096 bc0097 bc0098 bc0099 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0100 bc0101 bc0102 bc0103 bc0104 bc0105 bc0106 bc0107 bc0108 bc0109 bc0110 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0111 bc0112 bc0113 bc0114 bc0115 bc0116 bc0117 bc0118 bc0119 bc0120 bc0121 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0122 bc0123 bc0124 bc0125 bc0126 bc0127 bc0128 bc0129 bc0130 bc0131 bc0132 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0133 bc0134 bc0135 bc0136 bc0137 bc0138 bc0139 bc0140 bc0141 bc0142 bc0143 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0144 bc0145 bc0146 bc0147 bc0148 bc0149 bc0150 bc0151 bc0152 bc0153 bc0154 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0155 bc0156 bc0157 bc0158 bc0159 bc0160 bc0161 bc0162 bc0163 bc0164 bc0165 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0166 bc0167 bc0168 bc0169 bc0170 bc0171 bc0172 bc0173 bc0174 bc0175 bc0176 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0177 bc0178 bc0179 bc0180 bc0181 bc0182 bc0183 bc0184 bc0185 bc0186 bc0187 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0188 bc0189 bc0190 bc0191 bc0192 bc0193 bc0194 bc0195 bc0196 bc0197 bc0198 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0199 bc0200 bc0201 bc0202 bc0203 bc0204 bc0205 bc0206 bc0207 bc0208 bc0209 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0210 bc0211 bc0212 bc0213 bc0214 bc0215 bc0216 bc0217 bc0218 bc0219 bc0220 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0221 bc0222 bc0223 bc0224 bc0225 bc0226 bc0227 bc0228 bc0229 bc0230 bc0231 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0232 bc0233 bc0234 bc0235 bc0236 bc0237 bc0238 bc0239 bc0240 bc0241 bc0242 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0243 bc0244 bc0245 bc0246 bc0247 bc0248 bc0249 bc0250 bc0251 bc0252 bc0253 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0254 bc0255 bc0256 bc0257 bc0258 bc0259 bc0260 bc0261 bc0262 bc0263 bc0264 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0265 bc0266 bc0267 bc0268 bc0269 bc0270 bc0271 bc0272 bc0273 bc0274 bc0275 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0276 bc0277 bc0278 bc0279 bc0280 bc0281 bc0282 bc0283 bc0284 bc0285 bc0286 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0287 bc0288 bc0289 bc0290 bc0291 bc0292 bc0293 bc0294 bc0295 bc0296 bc0297 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0298 bc0299 bc0300 bc0301 bc0302 bc0303 bc0304 bc0305 bc0306 bc0307 bc0308 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0309 bc0310 bc0311 bc0312 bc0313 bc0314 bc0315 bc0316 bc0317 bc0318 bc0319 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0320 bc0321 bc0322 bc0323 bc0324 bc0325 bc0326 bc0327 bc0328 bc0329 bc0330 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0331 bc0332 bc0333 bc0334 bc0335 bc0336 bc0337 bc0338 bc0339 bc0340 bc0341 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0342 bc0343 bc0344 bc0345 bc0346 bc0347 bc0348 bc0349 bc0350 bc0351 bc0352 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0353 bc0354 bc0355 bc0356 bc0357 bc0358 bc0359 bc0360 bc0361 bc0362 bc0363 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0364 bc0365 bc0366 bc0367 bc0368 bc0369 bc0370 bc0371 bc0372 bc0373 bc0374 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0375 bc0376 bc0377 bc0378 bc0379 bc0380 bc0381 bc0382 bc0383 bc0384 bc0385 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0386 bc0387 bc0388 bc0389 bc0390 bc0391 bc0392 bc0393 bc0394 bc0395 bc0396 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0397 bc0398 bc0399 bc0400 bc0401 bc0402 bc0403 bc0404 bc0405 bc0406 bc0407 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0408 bc0409 bc0410 bc0411 bc0412 bc0413 bc0414 bc0415 bc0416 bc0417 bc0418 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0419 bc0420 bc0421 bc0422 bc0423 bc0424 bc0425 bc0426 bc0427 bc0428 bc0429 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0430 bc0431 bc0432 bc0433 bc0434 bc0435 bc0436 bc0437 bc0438 bc0439 bc0440 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0441 bc0442 bc0443 bc0444 bc0445 bc0446 bc0447 bc0448 bc0449 bc0450 bc0451 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0452 bc0453 bc0454 bc0455 bc0456 bc0457 bc0458 bc0459 bc0460 bc0461 bc0462 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0463 bc0464 bc0465 bc0466 bc0467 bc0468 bc0469 bc0470 bc0471 bc0472 bc0473 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0474 bc0475 bc0476 bc0477 bc0478 bc0479 bc0480 bc0481 bc0482 bc0483 bc0484 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc0485 bc0486 bc0487 bc0488 bc0489 bc0490 bc0491 bc0492 bc0493 bc0494 bc0495 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
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## bc3004 bc3005 bc3006 bc3007 bc3008 bc3009 bc3010 bc3011 bc3012 bc3013 bc3014 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3015 bc3016 bc3017 bc3018 bc3019 bc3020 bc3021 bc3022 bc3023 bc3024 bc3025 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3026 bc3027 bc3028 bc3029 bc3030 bc3031 bc3032 bc3033 bc3034 bc3035 bc3036 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3037 bc3038 bc3039 bc3040 bc3041 bc3042 bc3043 bc3044 bc3045 bc3046 bc3047 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3048 bc3049 bc3050 bc3051 bc3052 bc3053 bc3054 bc3055 bc3056 bc3057 bc3058 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3059 bc3060 bc3061 bc3062 bc3063 bc3064 bc3065 bc3066 bc3067 bc3068 bc3069 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3070 bc3071 bc3072 bc3073 bc3074 bc3075 bc3076 bc3077 bc3078 bc3079 bc3080 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3081 bc3082 bc3083 bc3084 bc3085 bc3086 bc3087 bc3088 bc3089 bc3090 bc3091 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3092 bc3093 bc3094 bc3095 bc3096 bc3097 bc3098 bc3099 bc3100 bc3101 bc3102 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3103 bc3104 bc3105 bc3106 bc3107 bc3108 bc3109 bc3110 bc3111 bc3112 bc3113 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3114 bc3115 bc3116 bc3117 bc3118 bc3119 bc3120 bc3121 bc3122 bc3123 bc3124 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3125 bc3126 bc3127 bc3128 bc3129 bc3130 bc3131 bc3132 bc3133 bc3134 bc3135 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3136 bc3137 bc3138 bc3139 bc3140 bc3141 bc3142 bc3143 bc3144 bc3145 bc3146 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3147 bc3148 bc3149 bc3150 bc3151 bc3152 bc3153 bc3154 bc3155 bc3156 bc3157 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3158 bc3159 bc3160 bc3161 bc3162 bc3163 bc3164 bc3165 bc3166 bc3167 bc3168 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3169 bc3170 bc3171 bc3172 bc3173 bc3174 bc3175 bc3176 bc3177 bc3178 bc3179 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3180 bc3181 bc3182 bc3183 bc3184 bc3185 bc3186 bc3187 bc3188 bc3189 bc3190 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3191 bc3192 bc3193 bc3194 bc3195 bc3196 bc3197 bc3198 bc3199 bc3200 bc3201 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3202 bc3203 bc3204 bc3205 bc3206 bc3207 bc3208 bc3209 bc3210 bc3211 bc3212 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3213 bc3214 bc3215 bc3216 bc3217 bc3218 bc3219 bc3220 bc3221 bc3222 bc3223 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3224 bc3225 bc3226 bc3227 bc3228 bc3229 bc3230 bc3231 bc3232 bc3233 bc3234 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3235 bc3236 bc3237 bc3238 bc3239 bc3240 bc3241 bc3242 bc3243 bc3244 bc3245 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3246 bc3247 bc3248 bc3249 bc3250 bc3251 bc3252 bc3253 bc3254 bc3255 bc3256 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3257 bc3258 bc3259 bc3260 bc3261 bc3262 bc3263 bc3264 bc3265 bc3266 bc3267 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3268 bc3269 bc3270 bc3271 bc3272 bc3273 bc3274 bc3275 bc3276 bc3277 bc3278 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3279 bc3280 bc3281 bc3282 bc3283 bc3284 bc3285 bc3286 bc3287 bc3288 bc3289 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3290 bc3291 bc3292 bc3293 bc3294 bc3295 bc3296 bc3297 bc3298 bc3299 bc3300 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3301 bc3302 bc3303 bc3304 bc3305 bc3306 bc3307 bc3308 bc3309 bc3310 bc3311 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3312 bc3313 bc3314 bc3315 bc3316 bc3317 bc3318 bc3319 bc3320 bc3321 bc3322 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3323 bc3324 bc3325 bc3326 bc3327 bc3328 bc3329 bc3330 bc3331 bc3332 bc3333 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3334 bc3335 bc3336 bc3337 bc3338 bc3339 bc3340 bc3341 bc3342 bc3343 bc3344 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3345 bc3346 bc3347 bc3348 bc3349 bc3350 bc3351 bc3352 bc3353 bc3354 bc3355 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3356 bc3357 bc3358 bc3359 bc3360 bc3361 bc3362 bc3363 bc3364 bc3365 bc3366 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3367 bc3368 bc3369 bc3370 bc3371 bc3372 bc3373 bc3374 bc3375 bc3376 bc3377 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3378 bc3379 bc3380 bc3381 bc3382 bc3383 bc3384 bc3385 bc3386 bc3387 bc3388 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3389 bc3390 bc3391 bc3392 bc3393 bc3394 bc3395 bc3396 bc3397 bc3398 bc3399 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3400 bc3401 bc3402 bc3403 bc3404 bc3405 bc3406 bc3407 bc3408 bc3409 bc3410 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3411 bc3412 bc3413 bc3414 bc3415 bc3416 bc3417 bc3418 bc3419 bc3420 bc3421 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3422 bc3423 bc3424 bc3425 bc3426 bc3427 bc3428 bc3429 bc3430 bc3431 bc3432 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3433 bc3434 bc3435 bc3436 bc3437 bc3438 bc3439 bc3440 bc3441 bc3442 bc3443 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3444 bc3445 bc3446 bc3447 bc3448 bc3449 bc3450 bc3451 bc3452 bc3453 bc3454 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3455 bc3456 bc3457 bc3458 bc3459 bc3460 bc3461 bc3462 bc3463 bc3464 bc3465 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3466 bc3467 bc3468 bc3469 bc3470 bc3471 bc3472 bc3473 bc3474 bc3475 bc3476 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3477 bc3478 bc3479 bc3480 bc3481 bc3482 bc3483 bc3484 bc3485 bc3486 bc3487 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3488 bc3489 bc3490 bc3491 bc3492 bc3493 bc3494 bc3495 bc3496 bc3497 bc3498 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3499 bc3500 bc3501 bc3502 bc3503 bc3504 bc3505 bc3506 bc3507 bc3508 bc3509 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3510 bc3511 bc3512 bc3513 bc3514 bc3515 bc3516 bc3517 bc3518 bc3519 bc3520 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3521 bc3522 bc3523 bc3524 bc3525 bc3526 bc3527 bc3528 bc3529 bc3530 bc3531 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3532 bc3533 bc3534 bc3535 bc3536 bc3537 bc3538 bc3539 bc3540 bc3541 bc3542 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3543 bc3544 bc3545 bc3546 bc3547 bc3548 bc3549 bc3550 bc3551 bc3552 bc3553 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3554 bc3555 bc3556 bc3557 bc3558 bc3559 bc3560 bc3561 bc3562 bc3563 bc3564 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3565 bc3566 bc3567 bc3568 bc3569 bc3570 bc3571 bc3572 bc3573 bc3574 bc3575 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3576 bc3577 bc3578 bc3579 bc3580 bc3581 bc3582 bc3583 bc3584 bc3585 bc3586 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3587 bc3588 bc3589 bc3590 bc3591 bc3592 bc3593 bc3594 bc3595 bc3596 bc3597 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3598 bc3599 bc3600 bc3601 bc3602 bc3603 bc3604 bc3605 bc3606 bc3607 bc3608 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3609 bc3610 bc3611 bc3612 bc3613 bc3614 bc3615 bc3616 bc3617 bc3618 bc3619 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3620 bc3621 bc3622 bc3623 bc3624 bc3625 bc3626 bc3627 bc3628 bc3629 bc3630 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3631 bc3632 bc3633 bc3634 bc3635 bc3636 bc3637 bc3638 bc3639 bc3640 bc3641 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3642 bc3643 bc3644 bc3645 bc3646 bc3647 bc3648 bc3649 bc3650 bc3651 bc3652 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3653 bc3654 bc3655 bc3656 bc3657 bc3658 bc3659 bc3660 bc3661 bc3662 bc3663 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3664 bc3665 bc3666 bc3667 bc3668 bc3669 bc3670 bc3671 bc3672 bc3673 bc3674 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3675 bc3676 bc3677 bc3678 bc3679 bc3680 bc3681 bc3682 bc3683 bc3684 bc3685 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3686 bc3687 bc3688 bc3689 bc3690 bc3691 bc3692 bc3693 bc3694 bc3695 bc3696 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3697 bc3698 bc3699 bc3700 bc3701 bc3702 bc3703 bc3704 bc3705 bc3706 bc3707 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3708 bc3709 bc3710 bc3711 bc3712 bc3713 bc3714 bc3715 bc3716 bc3717 bc3718 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3719 bc3720 bc3721 bc3722 bc3723 bc3724 bc3725 bc3726 bc3727 bc3728 bc3729 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3730 bc3731 bc3732 bc3733 bc3734 bc3735 bc3736 bc3737 bc3738 bc3739 bc3740 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3741 bc3742 bc3743 bc3744 bc3745 bc3746 bc3747 bc3748 bc3749 bc3750 bc3751 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3752 bc3753 bc3754 bc3755 bc3756 bc3757 bc3758 bc3759 bc3760 bc3761 bc3762 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3763 bc3764 bc3765 bc3766 bc3767 bc3768 bc3769 bc3770 bc3771 bc3772 bc3773 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3774 bc3775 bc3776 bc3777 bc3778 bc3779 bc3780 bc3781 bc3782 bc3783 bc3784 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3785 bc3786 bc3787 bc3788 bc3789 bc3790 bc3791 bc3792 bc3793 bc3794 bc3795 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3796 bc3797 bc3798 bc3799 bc3800 bc3801 bc3802 bc3803 bc3804 bc3805 bc3806 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3807 bc3808 bc3809 bc3810 bc3811 bc3812 bc3813 bc3814 bc3815 bc3816 bc3817 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3818 bc3819 bc3820 bc3821 bc3822 bc3823 bc3824 bc3825 bc3826 bc3827 bc3828 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## bc3829 bc3830 bc3831 bc3832 bc3833 bc3834 bc3835 bc3836 bc3837 bc3838 
##    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC    YuC 
## Levels: YuC
YuC$orig.ident <- "YuC"


YuT <- data.table::fread("/home/moyra.lawrence/R_analysis/Comparison/Yu/GSE166027_single_cell_gene_expression_CX5461(1).csv.gz", data.table = FALSE)
names(YuT)[2] <- 'Gene'
YuT <- YuT[,-1]
YuT <- YuT[,-2]
YuT <- YuT[,-2]
YuT <- YuT[,-2]
gene4 <- YuT$Gene
gene5 <- str_replace(gene4,
                    pattern = ".[0-9]+$",
                    replacement = "")
ens3 = getBM(attributes=c('ensembl_gene_id', 'mgi_symbol'), 
              filters = 'ensembl_gene_id', 
              values = gene5, 
              mart = ensembl)
YuT$Gene <- gene5

merge3 <- merge(YuT, ens2, by.x="Gene", by.y = "ensembl_gene_id")
merge3["mgi_symbol"][merge3["mgi_symbol"] == ''] <- "Unknown"
names3 <- make.unique(merge3$mgi_symbol)
names3 <- gsub("_", "-", names3)
names3 <- gsub("\\.", "-", names3)
names3 <- make.unique(names3)
rownames(merge3) <- names3
merge3 <- merge3[,-1] 
merge3 <- merge3[,-4923]

YuT <- CreateSeuratObject(counts=merge3, project="YuT")
YuT$orig.ident
## bc0001 bc0002 bc0003 bc0004 bc0005 bc0006 bc0007 bc0008 bc0009 bc0010 bc0011 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0012 bc0013 bc0014 bc0015 bc0016 bc0017 bc0018 bc0019 bc0020 bc0021 bc0022 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0023 bc0024 bc0025 bc0026 bc0027 bc0028 bc0029 bc0030 bc0031 bc0032 bc0033 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0034 bc0035 bc0036 bc0037 bc0038 bc0039 bc0040 bc0041 bc0042 bc0043 bc0044 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0045 bc0046 bc0047 bc0048 bc0049 bc0050 bc0051 bc0052 bc0053 bc0054 bc0055 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0056 bc0057 bc0058 bc0059 bc0060 bc0061 bc0062 bc0063 bc0064 bc0065 bc0066 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0067 bc0068 bc0069 bc0070 bc0071 bc0072 bc0073 bc0074 bc0075 bc0076 bc0077 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0078 bc0079 bc0080 bc0081 bc0082 bc0083 bc0084 bc0085 bc0086 bc0087 bc0088 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0089 bc0090 bc0091 bc0092 bc0093 bc0094 bc0095 bc0096 bc0097 bc0098 bc0099 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0100 bc0101 bc0102 bc0103 bc0104 bc0105 bc0106 bc0107 bc0108 bc0109 bc0110 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0111 bc0112 bc0113 bc0114 bc0115 bc0116 bc0117 bc0118 bc0119 bc0120 bc0121 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0122 bc0123 bc0124 bc0125 bc0126 bc0127 bc0128 bc0129 bc0130 bc0131 bc0132 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0133 bc0134 bc0135 bc0136 bc0137 bc0138 bc0139 bc0140 bc0141 bc0142 bc0143 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0144 bc0145 bc0146 bc0147 bc0148 bc0149 bc0150 bc0151 bc0152 bc0153 bc0154 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0155 bc0156 bc0157 bc0158 bc0159 bc0160 bc0161 bc0162 bc0163 bc0164 bc0165 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0166 bc0167 bc0168 bc0169 bc0170 bc0171 bc0172 bc0173 bc0174 bc0175 bc0176 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0177 bc0178 bc0179 bc0180 bc0181 bc0182 bc0183 bc0184 bc0185 bc0186 bc0187 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0188 bc0189 bc0190 bc0191 bc0192 bc0193 bc0194 bc0195 bc0196 bc0197 bc0198 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0199 bc0200 bc0201 bc0202 bc0203 bc0204 bc0205 bc0206 bc0207 bc0208 bc0209 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0210 bc0211 bc0212 bc0213 bc0214 bc0215 bc0216 bc0217 bc0218 bc0219 bc0220 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0221 bc0222 bc0223 bc0224 bc0225 bc0226 bc0227 bc0228 bc0229 bc0230 bc0231 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0232 bc0233 bc0234 bc0235 bc0236 bc0237 bc0238 bc0239 bc0240 bc0241 bc0242 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0243 bc0244 bc0245 bc0246 bc0247 bc0248 bc0249 bc0250 bc0251 bc0252 bc0253 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0254 bc0255 bc0256 bc0257 bc0258 bc0259 bc0260 bc0261 bc0262 bc0263 bc0264 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0265 bc0266 bc0267 bc0268 bc0269 bc0270 bc0271 bc0272 bc0273 bc0274 bc0275 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0276 bc0277 bc0278 bc0279 bc0280 bc0281 bc0282 bc0283 bc0284 bc0285 bc0286 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0287 bc0288 bc0289 bc0290 bc0291 bc0292 bc0293 bc0294 bc0295 bc0296 bc0297 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0298 bc0299 bc0300 bc0301 bc0302 bc0303 bc0304 bc0305 bc0306 bc0307 bc0308 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0309 bc0310 bc0311 bc0312 bc0313 bc0314 bc0315 bc0316 bc0317 bc0318 bc0319 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0320 bc0321 bc0322 bc0323 bc0324 bc0325 bc0326 bc0327 bc0328 bc0329 bc0330 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc0331 bc0332 bc0333 bc0334 bc0335 bc0336 bc0337 bc0338 bc0339 bc0340 bc0341 
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## bc4082 bc4083 bc4084 bc4085 bc4086 bc4087 bc4088 bc4089 bc4090 bc4091 bc4092 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4093 bc4094 bc4095 bc4096 bc4097 bc4098 bc4099 bc4100 bc4101 bc4102 bc4103 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4104 bc4105 bc4106 bc4107 bc4108 bc4109 bc4110 bc4111 bc4112 bc4113 bc4114 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4115 bc4116 bc4117 bc4118 bc4119 bc4120 bc4121 bc4122 bc4123 bc4124 bc4125 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4126 bc4127 bc4128 bc4129 bc4130 bc4131 bc4132 bc4133 bc4134 bc4135 bc4136 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4137 bc4138 bc4139 bc4140 bc4141 bc4142 bc4143 bc4144 bc4145 bc4146 bc4147 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4148 bc4149 bc4150 bc4151 bc4152 bc4153 bc4154 bc4155 bc4156 bc4157 bc4158 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4159 bc4160 bc4161 bc4162 bc4163 bc4164 bc4165 bc4166 bc4167 bc4168 bc4169 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4170 bc4171 bc4172 bc4173 bc4174 bc4175 bc4176 bc4177 bc4178 bc4179 bc4180 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4181 bc4182 bc4183 bc4184 bc4185 bc4186 bc4187 bc4188 bc4189 bc4190 bc4191 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4192 bc4193 bc4194 bc4195 bc4196 bc4197 bc4198 bc4199 bc4200 bc4201 bc4202 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4203 bc4204 bc4205 bc4206 bc4207 bc4208 bc4209 bc4210 bc4211 bc4212 bc4213 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4214 bc4215 bc4216 bc4217 bc4218 bc4219 bc4220 bc4221 bc4222 bc4223 bc4224 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4225 bc4226 bc4227 bc4228 bc4229 bc4230 bc4231 bc4232 bc4233 bc4234 bc4235 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4236 bc4237 bc4238 bc4239 bc4240 bc4241 bc4242 bc4243 bc4244 bc4245 bc4246 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4247 bc4248 bc4249 bc4250 bc4251 bc4252 bc4253 bc4254 bc4255 bc4256 bc4257 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4258 bc4259 bc4260 bc4261 bc4262 bc4263 bc4264 bc4265 bc4266 bc4267 bc4268 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4269 bc4270 bc4271 bc4272 bc4273 bc4274 bc4275 bc4276 bc4277 bc4278 bc4279 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4280 bc4281 bc4282 bc4283 bc4284 bc4285 bc4286 bc4287 bc4288 bc4289 bc4290 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4291 bc4292 bc4293 bc4294 bc4295 bc4296 bc4297 bc4298 bc4299 bc4300 bc4301 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4302 bc4303 bc4304 bc4305 bc4306 bc4307 bc4308 bc4309 bc4310 bc4311 bc4312 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4313 bc4314 bc4315 bc4316 bc4317 bc4318 bc4319 bc4320 bc4321 bc4322 bc4323 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4324 bc4325 bc4326 bc4327 bc4328 bc4329 bc4330 bc4331 bc4332 bc4333 bc4334 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4335 bc4336 bc4337 bc4338 bc4339 bc4340 bc4341 bc4342 bc4343 bc4344 bc4345 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4346 bc4347 bc4348 bc4349 bc4350 bc4351 bc4352 bc4353 bc4354 bc4355 bc4356 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4357 bc4358 bc4359 bc4360 bc4361 bc4362 bc4363 bc4364 bc4365 bc4366 bc4367 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4368 bc4369 bc4370 bc4371 bc4372 bc4373 bc4374 bc4375 bc4376 bc4377 bc4378 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4379 bc4380 bc4381 bc4382 bc4383 bc4384 bc4385 bc4386 bc4387 bc4388 bc4389 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4390 bc4391 bc4392 bc4393 bc4394 bc4395 bc4396 bc4397 bc4398 bc4399 bc4400 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4401 bc4402 bc4403 bc4404 bc4405 bc4406 bc4407 bc4408 bc4409 bc4410 bc4411 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4412 bc4413 bc4414 bc4415 bc4416 bc4417 bc4418 bc4419 bc4420 bc4421 bc4422 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4423 bc4424 bc4425 bc4426 bc4427 bc4428 bc4429 bc4430 bc4431 bc4432 bc4433 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4434 bc4435 bc4436 bc4437 bc4438 bc4439 bc4440 bc4441 bc4442 bc4443 bc4444 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4445 bc4446 bc4447 bc4448 bc4449 bc4450 bc4451 bc4452 bc4453 bc4454 bc4455 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4456 bc4457 bc4458 bc4459 bc4460 bc4461 bc4462 bc4463 bc4464 bc4465 bc4466 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4467 bc4468 bc4469 bc4470 bc4471 bc4472 bc4473 bc4474 bc4475 bc4476 bc4477 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4478 bc4479 bc4480 bc4481 bc4482 bc4483 bc4484 bc4485 bc4486 bc4487 bc4488 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4489 bc4490 bc4491 bc4492 bc4493 bc4494 bc4495 bc4496 bc4497 bc4498 bc4499 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4500 bc4501 bc4502 bc4503 bc4504 bc4505 bc4506 bc4507 bc4508 bc4509 bc4510 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4511 bc4512 bc4513 bc4514 bc4515 bc4516 bc4517 bc4518 bc4519 bc4520 bc4521 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4522 bc4523 bc4524 bc4525 bc4526 bc4527 bc4528 bc4529 bc4530 bc4531 bc4532 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4533 bc4534 bc4535 bc4536 bc4537 bc4538 bc4539 bc4540 bc4541 bc4542 bc4543 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4544 bc4545 bc4546 bc4547 bc4548 bc4549 bc4550 bc4551 bc4552 bc4553 bc4554 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4555 bc4556 bc4557 bc4558 bc4559 bc4560 bc4561 bc4562 bc4563 bc4564 bc4565 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4566 bc4567 bc4568 bc4569 bc4570 bc4571 bc4572 bc4573 bc4574 bc4575 bc4576 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4577 bc4578 bc4579 bc4580 bc4581 bc4582 bc4583 bc4584 bc4585 bc4586 bc4587 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4588 bc4589 bc4590 bc4591 bc4592 bc4593 bc4594 bc4595 bc4596 bc4597 bc4598 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4599 bc4600 bc4601 bc4602 bc4603 bc4604 bc4605 bc4606 bc4607 bc4608 bc4609 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4610 bc4611 bc4612 bc4613 bc4614 bc4615 bc4616 bc4617 bc4618 bc4619 bc4620 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4621 bc4622 bc4623 bc4624 bc4625 bc4626 bc4627 bc4628 bc4629 bc4630 bc4631 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4632 bc4633 bc4634 bc4635 bc4636 bc4637 bc4638 bc4639 bc4640 bc4641 bc4642 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4643 bc4644 bc4645 bc4646 bc4647 bc4648 bc4649 bc4650 bc4651 bc4652 bc4653 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4654 bc4655 bc4656 bc4657 bc4658 bc4659 bc4660 bc4661 bc4662 bc4663 bc4664 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4665 bc4666 bc4667 bc4668 bc4669 bc4670 bc4671 bc4672 bc4673 bc4674 bc4675 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4676 bc4677 bc4678 bc4679 bc4680 bc4681 bc4682 bc4683 bc4684 bc4685 bc4686 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4687 bc4688 bc4689 bc4690 bc4691 bc4692 bc4693 bc4694 bc4695 bc4696 bc4697 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4698 bc4699 bc4700 bc4701 bc4702 bc4703 bc4704 bc4705 bc4706 bc4707 bc4708 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4709 bc4710 bc4711 bc4712 bc4713 bc4714 bc4715 bc4716 bc4717 bc4718 bc4719 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4720 bc4721 bc4722 bc4723 bc4724 bc4725 bc4726 bc4727 bc4728 bc4729 bc4730 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4731 bc4732 bc4733 bc4734 bc4735 bc4736 bc4737 bc4738 bc4739 bc4740 bc4741 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4742 bc4743 bc4744 bc4745 bc4746 bc4747 bc4748 bc4749 bc4750 bc4751 bc4752 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4753 bc4754 bc4755 bc4756 bc4757 bc4758 bc4759 bc4760 bc4761 bc4762 bc4763 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4764 bc4765 bc4766 bc4767 bc4768 bc4769 bc4770 bc4771 bc4772 bc4773 bc4774 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4775 bc4776 bc4777 bc4778 bc4779 bc4780 bc4781 bc4782 bc4783 bc4784 bc4785 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4786 bc4787 bc4788 bc4789 bc4790 bc4791 bc4792 bc4793 bc4794 bc4795 bc4796 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4797 bc4798 bc4799 bc4800 bc4801 bc4802 bc4803 bc4804 bc4805 bc4806 bc4807 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4808 bc4809 bc4810 bc4811 bc4812 bc4813 bc4814 bc4815 bc4816 bc4817 bc4818 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4819 bc4820 bc4821 bc4822 bc4823 bc4824 bc4825 bc4826 bc4827 bc4828 bc4829 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4830 bc4831 bc4832 bc4833 bc4834 bc4835 bc4836 bc4837 bc4838 bc4839 bc4840 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4841 bc4842 bc4843 bc4844 bc4845 bc4846 bc4847 bc4848 bc4849 bc4850 bc4851 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4852 bc4853 bc4854 bc4855 bc4856 bc4857 bc4858 bc4859 bc4860 bc4861 bc4862 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4863 bc4864 bc4865 bc4866 bc4867 bc4868 bc4869 bc4870 bc4871 bc4872 bc4873 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4874 bc4875 bc4876 bc4877 bc4878 bc4879 bc4880 bc4881 bc4882 bc4883 bc4884 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4885 bc4886 bc4887 bc4888 bc4889 bc4890 bc4891 bc4892 bc4893 bc4894 bc4895 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4896 bc4897 bc4898 bc4899 bc4900 bc4901 bc4902 bc4903 bc4904 bc4905 bc4906 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4907 bc4908 bc4909 bc4910 bc4911 bc4912 bc4913 bc4914 bc4915 bc4916 bc4917 
##    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT    YuT 
## bc4918 bc4919 bc4920 bc4921 bc4922 
##    YuT    YuT    YuT    YuT    YuT 
## Levels: YuT
YuT$orig.ident <- "YuT"

#Plotting Yu

YuC[["percent.mt"]] <- PercentageFeatureSet(YuC, pattern = "^mt-")
YuT[["percent.mt"]] <- PercentageFeatureSet(YuT, pattern = "^mt-")
VlnPlot(YuC, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(YuT, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

##Filtering Yu 2000 < nFeature_RNA
5,000 < nCount_RNA <100,000
0 =< percent.mt < 10

YuC.filtered <- subset(YuC, subset = nFeature_RNA > 2000 & nCount_RNA > 5000 & nCount_RNA < 100000 & percent.mt < 10)
VlnPlot(YuC.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt" ), ncol = 3, log=TRUE)
YuT.filtered <- subset(YuT, subset = nFeature_RNA > 2000 & nCount_RNA > 5000 & nCount_RNA < 100000 & percent.mt < 10)
VlnPlot(YuT.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt" ), ncol = 3, log=TRUE)

#Shen

ShenNC<-read.delim(file = "/home/moyra.lawrence/R_analysis/Comparison/Shen/GSM5195022_SC-siNC-count.csv.gz", header = TRUE, sep = ",") 
ShenSnrpd2<-read.delim(file = "/home/moyra.lawrence/R_analysis/Comparison/Shen/GSM5195023_SC-siSnrpd2-count.csv.gz", header = TRUE, sep = ",") 
ShenPSC<-read.delim(file = "/home/moyra.lawrence/R_analysis/Comparison/Shen/GSM5195024_SC-PSC-count.csv.gz", header = TRUE, sep = ",") 
ShenTBLC<-read.delim(file = "/home/moyra.lawrence/R_analysis/Comparison/Shen/GSM5195025_SC-TBLC-count.csv.gz", header = TRUE, sep = ",") 
ShenNCS <- CreateSeuratObject(counts = ShenNC, project = "ShenNC", min.cells = 3, min.features = 200)
ShenSnrpd2S <- CreateSeuratObject(counts = ShenSnrpd2, project = "ShenSnrpd2", min.cells = 3, min.features = 200)
ShenPSCS <- CreateSeuratObject(counts = ShenPSC, project = "ShenPSC", min.cells = 3, min.features = 200)
ShenTBLCS <- CreateSeuratObject(counts = ShenTBLC, project = "ShenTBLC", min.cells = 3, min.features = 200)

#Plotting Shen

ShenNCS[["percent.mt"]] <- PercentageFeatureSet(ShenNCS, pattern = "^mt-")
ShenSnrpd2S[["percent.mt"]] <- PercentageFeatureSet(ShenSnrpd2S, pattern = "^mt-")
ShenPSCS[["percent.mt"]] <- PercentageFeatureSet(ShenPSCS, pattern = "^mt-")
ShenTBLCS[["percent.mt"]] <- PercentageFeatureSet(ShenTBLCS, pattern = "^mt-")
VlnPlot(ShenNCS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(ShenSnrpd2S, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(ShenPSCS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

VlnPlot(ShenTBLCS, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3, log=TRUE)

Loading embryo data and preparing for seurat

raw_data <- data.table::fread("/home/moyra.lawrence/R_analysis/Second_10x_run/GSE45719_matrix.txt.gz", data.table = FALSE)

names(raw_data)[1] <- 'Gene'
names <- make.unique(raw_data$Gene)
rownames(raw_data) <- names
raw_data <- raw_data[,-1] # get rid of old names

colnames(raw_data) <- gsub("_", "-", colnames(raw_data))
colnames(raw_data) <- stri_replace_all_fixed(str = colnames(raw_data), pattern = "_", replacement = "-")
colnames(raw_data) <- stri_replace_all_fixed(str = colnames(raw_data), pattern = "zy1", replacement = "zygote-1")
colnames(raw_data) <- stri_replace_all_fixed(str = colnames(raw_data), pattern = "zy2", replacement = "zygote-2")
colnames(raw_data) <- stri_replace_all_fixed(str = colnames(raw_data), pattern = "zy3", replacement = "zygote-3")
colnames(raw_data) <- stri_replace_all_fixed(str = colnames(raw_data), pattern = "zy4", replacement = "zygote-4")

ML11 <- CreateSeuratObject(counts=raw_data, project="ML11", names.delim="-")
## Warning: Feature names cannot have pipe characters ('|'), replacing with dashes
## ('-')
ML11$orig.ident
##            16cell-1-10            16cell-1-11            16cell-1-12 
##                 16cell                 16cell                 16cell 
##            16cell-1-13            16cell-1-14            16cell-1-15 
##                 16cell                 16cell                 16cell 
##             16cell-1-2             16cell-1-3             16cell-1-4 
##                 16cell                 16cell                 16cell 
##             16cell-1-5             16cell-1-6             16cell-1-7 
##                 16cell                 16cell                 16cell 
##             16cell-1-8             16cell-1-9             16cell-4-1 
##                 16cell                 16cell                 16cell 
##            16cell-4-10            16cell-4-11             16cell-4-2 
##                 16cell                 16cell                 16cell 
##             16cell-4-3             16cell-4-4             16cell-4-5 
##                 16cell                 16cell                 16cell 
##             16cell-4-6             16cell-4-7             16cell-4-8 
##                 16cell                 16cell                 16cell 
##             16cell-4-9             16cell-5-1            16cell-5-10 
##                 16cell                 16cell                 16cell 
##            16cell-5-11            16cell-5-12            16cell-5-13 
##                 16cell                 16cell                 16cell 
##             16cell-5-2             16cell-5-3             16cell-5-4 
##                 16cell                 16cell                 16cell 
##             16cell-5-5             16cell-5-6             16cell-5-7 
##                 16cell                 16cell                 16cell 
##             16cell-5-8             16cell-5-9             16cell-6-1 
##                 16cell                 16cell                 16cell 
##            16cell-6-10            16cell-6-11            16cell-6-12 
##                 16cell                 16cell                 16cell 
##             16cell-6-2             16cell-6-3             16cell-6-4 
##                 16cell                 16cell                 16cell 
##             16cell-6-5             16cell-6-6             16cell-6-7 
##                 16cell                 16cell                 16cell 
##             16cell-6-8             16cell-6-9              4cell-1-1 
##                 16cell                 16cell                  4cell 
##              4cell-1-2              4cell-1-4              4cell-2-1 
##                  4cell                  4cell                  4cell 
##              4cell-2-2              4cell-2-3              4cell-2-4 
##                  4cell                  4cell                  4cell 
##              4cell-3-1              4cell-3-3              4cell-3-4 
##                  4cell                  4cell                  4cell 
##              4cell-4-1              4cell-4-2              4cell-4-3 
##                  4cell                  4cell                  4cell 
##              4cell-4-4              8cell-1-1              8cell-1-2 
##                  4cell                  8cell                  8cell 
##              8cell-1-4              8cell-1-5              8cell-1-6 
##                  8cell                  8cell                  8cell 
##              8cell-1-7              8cell-1-8              8cell-2-1 
##                  8cell                  8cell                  8cell 
##              8cell-2-2              8cell-2-3              8cell-2-4 
##                  8cell                  8cell                  8cell 
##              8cell-2-6              8cell-2-7              8cell-2-8 
##                  8cell                  8cell                  8cell 
##              8cell-5-1              8cell-5-2              8cell-5-3 
##                  8cell                  8cell                  8cell 
##              8cell-5-4              8cell-5-6              8cell-5-7 
##                  8cell                  8cell                  8cell 
##              8cell-5-8              8cell-8-1              8cell-8-2 
##                  8cell                  8cell                  8cell 
##              8cell-8-3              8cell-8-4              8cell-8-6 
##                  8cell                  8cell                  8cell 
##              8cell-8-7              8cell-8-8  BXC-100pg-liver-RNA-1 
##                  8cell                  8cell                    BXC 
##  BXC-100pg-liver-RNA-2   BXC-10pg-liver-RNA-1   BXC-10pg-liver-RNA-2 
##                    BXC                    BXC                    BXC 
##   BXC-1ng-liver-RNA-0r    BXC-1ng-liver-RNA-1  BXC-30pg-liver-RNA-0r 
##                    BXC                    BXC                    BXC 
##   BXC-30pg-liver-RNA-2       BXC-liver-cell-1       BXC-liver-cell-2 
##                    BXC                    BXC                    BXC 
##       BXC-liver-cell-4       BXC-liver-cell-5       BXC-liver-cell-6 
##                    BXC                    BXC                    BXC 
##        C57twocell-16-1        C57twocell-16-2        C57twocell-18-1 
##             C57twocell             C57twocell             C57twocell 
##        C57twocell-18-2        C57twocell-20-1        C57twocell-20-2 
##             C57twocell             C57twocell             C57twocell 
##        C57twocell-22-1        C57twocell-22-2        early2cell-0r-1 
##             C57twocell             C57twocell             early2cell 
##        early2cell-0r-2         early2cell-1-1         early2cell-1-2 
##             early2cell             early2cell             early2cell 
##         early2cell-2-1         early2cell-2-2         early2cell-3-1 
##             early2cell             early2cell             early2cell 
##         early2cell-3-2         earlyblast-2-1        earlyblast-2-10 
##             early2cell             earlyblast             earlyblast 
##        earlyblast-2-12        earlyblast-2-15        earlyblast-2-16 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-2-17        earlyblast-2-19         earlyblast-2-2 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-2-22         earlyblast-2-3         earlyblast-2-4 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-2-5         earlyblast-2-6         earlyblast-2-7 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-2-8         earlyblast-3-1        earlyblast-3-10 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-3-12        earlyblast-3-13        earlyblast-3-14 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-3-15        earlyblast-3-16        earlyblast-3-17 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-3-2         earlyblast-3-3         earlyblast-3-4 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-3-6         earlyblast-3-7         earlyblast-3-8 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-3-9         earlyblast-4-1        earlyblast-4-12 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-4-13        earlyblast-4-14        earlyblast-4-16 
##             earlyblast             earlyblast             earlyblast 
##        earlyblast-4-17        earlyblast-4-18         earlyblast-4-2 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-4-3         earlyblast-4-5         earlyblast-4-6 
##             earlyblast             earlyblast             earlyblast 
##         earlyblast-4-8         earlyblast-4-9          late2cell-5-1 
##             earlyblast             earlyblast              late2cell 
##          late2cell-5-2          late2cell-6-1          late2cell-6-2 
##              late2cell              late2cell              late2cell 
##          late2cell-7-1          late2cell-7-2          late2cell-8-1 
##              late2cell              late2cell              late2cell 
##          late2cell-8-2          late2cell-9-1          late2cell-9-2 
##              late2cell              late2cell              late2cell 
##         lateblast-1-10         lateblast-1-11         lateblast-1-13 
##              lateblast              lateblast              lateblast 
##         lateblast-1-14         lateblast-1-16         lateblast-1-19 
##              lateblast              lateblast              lateblast 
##          lateblast-1-2         lateblast-1-20         lateblast-1-21 
##              lateblast              lateblast              lateblast 
##         lateblast-1-23         lateblast-1-24         lateblast-1-26 
##              lateblast              lateblast              lateblast 
##         lateblast-1-27          lateblast-1-4          lateblast-1-5 
##              lateblast              lateblast              lateblast 
##          lateblast-1-6          lateblast-1-7          lateblast-1-8 
##              lateblast              lateblast              lateblast 
##          lateblast-1-9          lateblast-2-1         lateblast-2-12 
##              lateblast              lateblast              lateblast 
##         lateblast-2-14         lateblast-2-16         lateblast-2-17 
##              lateblast              lateblast              lateblast 
##          lateblast-2-2          lateblast-2-3          lateblast-2-5 
##              lateblast              lateblast              lateblast 
##          lateblast-2-7          lateblast-2-8          lateblast-2-9 
##              lateblast              lateblast              lateblast 
##          mid2cell-0r-1          mid2cell-0r-2           mid2cell-3-1 
##               mid2cell               mid2cell               mid2cell 
##           mid2cell-3-2           mid2cell-4-1           mid2cell-4-2 
##               mid2cell               mid2cell               mid2cell 
##           mid2cell-5-1           mid2cell-5-2           mid2cell-6-1 
##               mid2cell               mid2cell               mid2cell 
##           mid2cell-6-2           mid2cell-7-1           mid2cell-7-2 
##               mid2cell               mid2cell               mid2cell 
##           midblast-1-1          midblast-1-10          midblast-1-11 
##               midblast               midblast               midblast 
##          midblast-1-12          midblast-1-13          midblast-1-14 
##               midblast               midblast               midblast 
##          midblast-1-15          midblast-1-16          midblast-1-17 
##               midblast               midblast               midblast 
##          midblast-1-18          midblast-1-19           midblast-1-2 
##               midblast               midblast               midblast 
##          midblast-1-20          midblast-1-22          midblast-1-23 
##               midblast               midblast               midblast 
##          midblast-1-24           midblast-1-3           midblast-1-4 
##               midblast               midblast               midblast 
##           midblast-1-5           midblast-1-6           midblast-1-8 
##               midblast               midblast               midblast 
##           midblast-1-9           midblast-2-1          midblast-2-10 
##               midblast               midblast               midblast 
##          midblast-2-11          midblast-2-12          midblast-2-13 
##               midblast               midblast               midblast 
##          midblast-2-14          midblast-2-15          midblast-2-16 
##               midblast               midblast               midblast 
##          midblast-2-17          midblast-2-18           midblast-2-2 
##               midblast               midblast               midblast 
##          midblast-2-23          midblast-2-24           midblast-2-3 
##               midblast               midblast               midblast 
##           midblast-2-4           midblast-2-5           midblast-2-6 
##               midblast               midblast               midblast 
##           midblast-2-7           midblast-2-8           midblast-2-9 
##               midblast               midblast               midblast 
##           midblast-3-1          midblast-3-10          midblast-3-11 
##               midblast               midblast               midblast 
##          midblast-3-12          midblast-3-13          midblast-3-14 
##               midblast               midblast               midblast 
##          midblast-3-15          midblast-3-17          midblast-3-18 
##               midblast               midblast               midblast 
##           midblast-3-2          midblast-3-23           midblast-3-3 
##               midblast               midblast               midblast 
##           midblast-3-4           midblast-3-5           midblast-3-6 
##               midblast               midblast               midblast 
##           midblast-3-7           midblast-3-8           midblast-3-9 
##               midblast               midblast               midblast 
##               zygote-1               zygote-2               zygote-3 
##                 zygote                 zygote                 zygote 
##               zygote-4 16cell-2pooled-split6a 16cell-2pooled-split6b 
##                 zygote                 16cell                 16cell 
## 16cell-2pooled-split8a 16cell-2pooled-split8b         16cell-split1a 
##                 16cell                 16cell                 16cell 
##         16cell-split1b         16cell-split2a         16cell-split2b 
##                 16cell                 16cell                 16cell 
##   8cell-12-1-smartseq2   8cell-12-2-smartseq2   8cell-12-3-smartseq2 
##                  8cell                  8cell                  8cell 
##   8cell-12-4-smartseq2   8cell-13-1-smartseq2   8cell-14-1-smartseq2 
##                  8cell                  8cell                  8cell 
##   8cell-14-2-smartseq2   8cell-14-3-smartseq2   8cell-14-4-smartseq2 
##                  8cell                  8cell                  8cell 
##  8cell-2pooled-split5a  8cell-2pooled-split5b  8cell-2pooled-split7a 
##                  8cell                  8cell                  8cell 
##  8cell-2pooled-split7b  8cell-2pooled-split9a  8cell-2pooled-split9b 
##                  8cell                  8cell                  8cell 
##          8cell-split3a          8cell-split3b          8cell-split4a 
##                  8cell                  8cell                  8cell 
##          8cell-split4b      fibroblast-13-CxB      fibroblast-14-CxB 
##                  8cell             fibroblast             fibroblast 
##      fibroblast-15-CxB      fibroblast-16-CxB      fibroblast-17-BxC 
##             fibroblast             fibroblast             fibroblast 
##      fibroblast-19-BxC      fibroblast-20-BxC      fibroblast-21-BxC 
##             fibroblast             fibroblast             fibroblast 
##      fibroblast-22-BxC       fibroblast-9-CxB 
##             fibroblast             fibroblast 
## 13 Levels: 16cell 4cell 8cell BXC C57twocell early2cell ... zygote
ML11[["in.vivo"]] <- Idents(object = ML11)
ML11$orig.ident <- "ML11"

Loading data

ML12 <- read_rds(file='/home/moyra.lawrence/count3/enumerate/EVEV/seurat.obj.rds')
ML13 <- read_rds(file='/home/moyra.lawrence/count3/enumerate/Dppa2_MERVL_TT/seurat.obj.rds')

ML12
## An object of class Seurat 
## 32294 features across 3438 samples within 1 assay 
## Active assay: RNA (32294 features, 0 variable features)
ML13
## An object of class Seurat 
## 32294 features across 3958 samples within 1 assay 
## Active assay: RNA (32294 features, 0 variable features)

##Filtering

ML12[["percent.mt"]] <- PercentageFeatureSet(ML12, pattern = "^mt-")
ML13[["percent.mt"]] <- PercentageFeatureSet(ML13, pattern = "^mt-")

Filtering conditions for EVEV

3500 < nFeature_RNA
10,000 < nCount_RNA <120,000
0 =< percent.mt < 10

Filtering conditions for DMTT

2000 < nFeature_RNA
10,000 < nCount_RNA <120,000
0 =< percent.mt < 15

Filtering datasets

ML12.filtered <- subset(ML12, subset = nFeature_RNA > 3500 & nCount_RNA > 10000 & nCount_RNA < 120000 & percent.mt < 10)
ML13.filtered <- subset(ML13, subset = nFeature_RNA > 2000 & nCount_RNA > 10000 & nCount_RNA < 120000 & percent.mt < 15)
VlnPlot(ML12.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt" ), ncol = 3, log=TRUE)

VlnPlot(ML13.filtered, pt.size=0, features = c("nFeature_RNA", "nCount_RNA", "percent.mt" ), ncol = 3, log=TRUE)

#Fixing orig.ident

Hu.filtered$orig.ident <- "Hu"
XuESC1.filtered$orig.ident <- "XuESC1"
XuESC2.filtered$orig.ident <- "XuESC2"
XuEPS1.filtered$orig.ident <- "XuEPS1"
XuEPS2.filtered$orig.ident <- "XuEPS2"
XuEPSTPS.filtered$orig.ident <- "XuEPSTPS"
Xu2CTPS.filtered$orig.ident <- "Xu2CTPS"
ShenNCS$orig.ident <- "ShenNCS"
ShenSnrpd2S$orig.ident <- "ShenSnrpd2S"
ShenPSCS$orig.ident <- "ShenPSCS"
ShenTBLCS$orig.ident <- "ShenTBLCS"

Merging data

merge.filtered <- merge(ML12.filtered, y = c(ML11, ML13.filtered, Hu.filtered, XuESC1.filtered, XuESC2.filtered, XuEPS1.filtered, XuEPS2.filtered, XuEPSTPS.filtered, Xu2CTPS.filtered, Cossec.filtered, YuC.filtered, YuT.filtered, ShenNCS, ShenSnrpd2S, ShenPSCS, ShenTBLCS), add.cell.ids = c("Empty_puro_Empty_hygro", "Embryo", "Dppa2_MERVL_TT", "Hu", "XuESC1", "XuESC2", "XuEPS1", "XuEPS2", "XuEPSTPS", "Xu2CTPS", "Cossec", "Yu_Control", "Yu_CX5461", "ShenNC", "ShenSnrpd2", "ShenPSC", "ShenTBLC"), project = "ML1")
merge.filtered
## An object of class Seurat 
## 59223 features across 92170 samples within 1 assay 
## Active assay: RNA (59223 features, 0 variable features)
save(merge.filtered, file="merge_filtered3.rda")
#load(file='merge_filtered3.rda')